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Journal of Experimental Biology, Vol 203, Issue 12 1835-1844, Copyright © 2000 by Company of Biologists

JOURNAL ARTICLES

Ion channel activity from the midgut brush-border membrane of gypsy moth (Lymantria dispar) larvae

O Peyronnet, V Vachon, JL Schwartz and R Laprade
Groupe de Recherche en Transport Membranaire, Universite de Montreal, Quebec, Canada.

Ion channels from the midgut apical membrane of gypsy moth (Lymantria dispar) larvae were studied following mechanical fusion of brush-border membrane vesicles with planar phospholipid bilayer membranes. In symmetrical 300 mmol l(-)(1) KCl (pH 9.0), nine different channels with conductances ranging from 27 to 795 pS and linear current/voltage relationships were resolved. In the presence of a KCl gradient across the bilayer (450 mmol l(-)(1 )cis/150 mmol l(-)(1 )trans), 11 different conductance levels ranging from 16 to 850 pS were detected. The channels were slightly cationic: the zero-current reversal potential was shifted by -5 mV to -21 mV compared with symmetrical KCl conditions, corresponding to p(K)/p(Cl) permeability ratios of 1.5-8.0. Most channels were neither voltage-dependent nor Ca(2+)-sensitive and displayed complex gating kinetics. Addition of Ba(2+) or Cs(+) to both sides of the bilayer had little effect on channel activity, but fewer distinct channels were observed when KCl was replaced by potassium gluconate, suggesting an effect of Cl(-) on channel activity. A reduced number of channels was also detected when KCl was replaced by N-methyl-d-glucamine-HCl. Under asymmetrical N-methyl-d-glucamine-HCl conditions, only anionic channels were observed. They exhibited current rectification (35 pS at negative voltages and 81 pS at positive voltages) and were strongly voltage-dependent.




© The Company of Biologists Ltd 2000